3-Hydroxyolean-12-en-27-oic Acids Inhibit RANKL-Induced Osteoclastogenesis in Vitro and Inflammation-Induced Bone Loss in Vivo
Abstract
:1. Introduction
2. Results
2.1. AR-3 and AR-5 Inhibits RANKL-Induced Osteoclast Formation in RAW264.7 Cells
2.2. AR-3 and AR-5 Inhibit RANKL-Induced Osteoclast Formation in Mouse BMMs
2.3. AR-3 and AR-5 Suppress RANKL-Induced Formation of Mature Osteoclasts in Mouse BMMs
2.4. AR-3 and AR-5 Suppress LPS-Induced Bone Loss in Vivo
2.5. AR-3 and AR-5 Suppress RANKL-Induced NFATc1 Activation
2.6. AR-5 Inhibits RANKL-Induced Osteoclast Formation at Early and Late Stage of Differentiation, and Migration
2.7. AR-5 Inhibits RANKL-Induced Activation of ERK and JNK MAPKs
3. Discussion
4. Materials and Methods
4.1. Chemicals, Reagents, and Antibodies
4.2. Cell Culture
4.3. In Vitro Osteoclastogenesis Assay
4.4. Measurement of Cell Viability
4.5. Bone Resorption Assay
4.6. Actin-Ring Formation Assay
4.7. Migration Assay
4.8. Western Blot Analysis
4.9. Immunofluorescence and Confocal Microscopy
4.10. Reverse Transcription qPCR Analysis
4.11. Animal Experiments
4.12. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
Abbreviations
AR-1 | 3α,23-isopropylidenedioxyolean-12-en-27-oic acid |
AR-2 | 3-oxoolean-12-en-27-oic acid |
AR-3 | 3α-hydroxyolean-12-en-27-oic acid |
AR-4 | 23-hydroxy-3-oxoolean-12-en-27-oic acid |
AR-5 | aceriphyllic acid A |
BMM | bone marrow-derived macrophage |
BS/BV | bone surface/bone volume |
CT | Computed Tomography |
CtsK | cathepsin K |
DAPI | 4′,6-diamidino-2-phenylindole |
DC-STAMP | dendritic cell–specific transmembrane protein |
ERK | extracellular signal-regulated kinase |
FITC | fluorescein isothiocyanate |
JNK | c-Jun N-terminal kinase |
LPS | lipopolysaccharide |
MAPK | mitogen-activated protein kinase |
M-CSF | macrophage colony-stimulating factor |
MMP-9 | matrix metallopeptidase 9 |
MTT | 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide |
NFATc1 | nuclear factor of activated T cells cytoplasmic 1 |
NF-κB | nuclear factor-κB |
OSCAR | osteoclast-associated receptor |
qPCR | quantitative polymerase chain reaction |
RANK | receptor activator of nuclear factor-κB |
RANKL | receptor activator of nuclear factor-κB ligand |
Tb.N | trabecular number |
Tb.Sp | trabecular separation |
TARF6 | TNF-receptor-associated factor 6 |
TRAP | tartarate-resistant acid phosphatase |
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Seo, W.; Lee, S.; Tran, P.T.; Ngo, T.Q.-M.; Kim, O.; Le, T.H.; Dang, N.H.; Hwangbo, C.; Min, B.S.; Lee, J.-H. 3-Hydroxyolean-12-en-27-oic Acids Inhibit RANKL-Induced Osteoclastogenesis in Vitro and Inflammation-Induced Bone Loss in Vivo. Int. J. Mol. Sci. 2020, 21, 5240. https://doi.org/10.3390/ijms21155240
Seo W, Lee S, Tran PT, Ngo TQ-M, Kim O, Le TH, Dang NH, Hwangbo C, Min BS, Lee J-H. 3-Hydroxyolean-12-en-27-oic Acids Inhibit RANKL-Induced Osteoclastogenesis in Vitro and Inflammation-Induced Bone Loss in Vivo. International Journal of Molecular Sciences. 2020; 21(15):5240. https://doi.org/10.3390/ijms21155240
Chicago/Turabian StyleSeo, Wonyoung, Suhyun Lee, Phuong Thao Tran, Thi Quynh-Mai Ngo, Okwha Kim, Thanh Huong Le, Nguyen Hai Dang, Cheol Hwangbo, Byung Sun Min, and Jeong-Hyung Lee. 2020. "3-Hydroxyolean-12-en-27-oic Acids Inhibit RANKL-Induced Osteoclastogenesis in Vitro and Inflammation-Induced Bone Loss in Vivo" International Journal of Molecular Sciences 21, no. 15: 5240. https://doi.org/10.3390/ijms21155240